The objective of the research is to define possible controls for DNA biosynthesis during normal and abnormal growth. Most of the DNA polymerase Alpha activity from HeLa cells in S-phase has been isolated as a Mr 640,000 multiprotein form that is designated DNA polymerase Alpha2. The enzyme has been purified to electrophoretic homogeneity and resolved into its components that include; a Mr 180,000 polymerase Alpha subunit, a Mr 70,000 DNA primase, a Mr 69,000 3'---greater than 5' exonuclease, Mr 96,000 C1 and Mr 52,000 C2 primer-recognition proteins and Mr 92,000 Ap A binding protein. The interaction of DNA polymerase Alpha2 and other required proteins for the efficient and specific initiation of RNA primer synthesis will be defined. The conditions and protein requirements in addition to polymerase Alpha2 that are required for elongation of DNA chains with catalytic efficiency, processivity and fidelity will be ascertained. The enzymic and nonenzymic proteins in addition to DNA polymerase Alpha2 that are required for the replicationof SV40-DNA in vitro in the presence of T-antigen will be studied.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA015187-15
Application #
3164120
Study Section
Biochemistry Study Section (BIO)
Project Start
1978-04-01
Project End
1991-11-30
Budget Start
1990-12-01
Budget End
1991-11-30
Support Year
15
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Worcester Foundation for Biomedical Research
Department
Type
DUNS #
City
Shrewsbury
State
MA
Country
United States
Zip Code
01545
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SenGupta, D N; Kumar, P; Zmudzka, B Z et al. (1987) Mammalian alpha-polymerase: cloning of partial complementary DNA and immunobinding of catalytic subunit in crude homogenate protein blots. Biochemistry 26:956-63

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